Cloning and expression of circular transcript of mouse growth hormone receptor gene.

Based on reports in the literature and search results on the circBase database, 8 circular transcripts of the mouse growth hormone receptor (GHR) gene were identified. In order to confirm the existence of the circular transcripts of the GHR gene (circGHRs) and to explore their expression patterns, the Kunming mouse (Mus musculus) was used as a research animal. This study detected the existence of circGHRs by RT-PCR amplification and sequencing, one of which was selected as circGHR for detailed analysis. The circular structure of circGHR was confirmed by RNase R treatment and reverse transcription. The spatiotemporal expression of circGHR and GHR mRNA was analyzed by qRT-PCR. The results showed that the full length of mouse circGHR was 820 nt, which was formed by circularization of exons 2-8 of the transcript of the GHR gene. RNase R tolerance analysis shows that mouse circGHR has the general characteristics of circular molecules and is not easily degraded by RNase R. Compared with oligo-d(T)18 primers, random primers have higher reverse transcription efficiency for circGHR, which further shows that circGHR is a poly(A)-free cyclic structure molecule. Tissue expression profile results show that circGHR is highly expressed in the liver and kidney of 1 week-old and 7-week old Kunming mice, but is low in pectoral muscles and leg muscles. The time-series expression profile of circGHR does not show any significant difference between the liver and pectoral muscle tissue. The circGHR expression in the leg muscle was low before 5 weeks of age but increased after 7 weeks of age. This study confirmed the existence of a circular transcript circGHR of the mouse GHR gene, and initially revealed the expression pattern of circGHR. The results of the study laid a foundation for in-depth developmental studies on the biological functions of the mouse circGHR and its mechanism of action regarding the growth and development of mice.

[Cloning and expression analysis of chicken circular transcript of insulin degrading enzyme gene].

Insulin-degrading enzyme (IDE) is a highly conserved metallopeptidase that functions in the catabolism of bioactive peptides. In our previous study, we identified a putative circular transcript in that chicken insulin-degrading enzyme (IDE) gene through analyzing a high throughput sequencing result. Here we set to confirm the circular transcript of IDE (circIDE) and explore its expression regularity in normal barred Plymouth chicken. The circIDE was confirmed by PCR amplification and sequencing. The circular structure of circIDE was determined by RNase R processing and reverse transcription experiments. Then we analyzed the spatiotemporal expression pattern of circIDE and IDE mRNA and compared the differential expression of circIDE and IDE mRNA in the normal barred Plymouth chicken and the dwarf ones. The results showed that the full length of chicken circIDE was 1332 nt, divided form exon 2-11 of the IDE gene. RNase R tolerance analysis showed that chicken circIDE had the general characteristics of circular molecule, and was highly resistant to RNase R. The random primers had higher transcription efficiency than the oligo-d(T)18 primers, confirming that circIDE is a circular structured molecule without poly(A). circIDE was highly expressed in the liver and heart tissues but less in the muscle tissues of leg and breast in normal chickens at the age of 1 and 12 weeks. The expression profile of circIDE in liver tissue showed that circIDE level was lower in1 to 6 weeks and then became higher after 8 weeks of age. The expression of circIDE in liver tissue was significantly higher in normal chicken than that in dwarf barred Plymouth chicken (P<0.05). This study confirmed a circIDE strucutre in chicken IDE gene and uncovered its expression regularity. We demonstrated that the expression level of circIDE in the liver tissue was higher in normal barred Plymouth chicken compared to dwarf species. This study paves the way for further understanding the biological function of chicken circIDE, including its roles in regulating chicken growth and development.

Optimised extraction of ß-carotene from Spirulina platensis and hypoglycaemic effect in streptozotocin-induced diabetic mice.

BACKGROUND: Spirulina platensis is rich in ß-carotene, which possesses many important biological activities. This study investigated the ultrasound-assisted extraction and purification of ß-carotene from Spirulina platensis by using response surface methodology (RSM), determined its antioxidant capacity in vitro and explored its hypoglycaemic effect in diabetic mice. RESULTS: The raw ß-carotene extract with a concentration of 1942.14 ± 10.03 µg mL(-1) was obtained at the optimised condition by RSM (0.40 of the solid-liquid ratio, 51% of the extraction power, and 17 min of the extraction time), and the purity of evaporated ß-carotene extract reached 816.32 ± 10.57 mg g(-1) after purified by a NKA-9 resin with a sampling and elution rate of 1 mL min(-1) . The ß-carotene extract scavenged 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radicals with the highest ratios of 44 ± 0.26% and 35 ± 0.45% respectively, and exhibited strong inhibiting capacity on anti-lipid peroxidation. The blood glucose level of streptozotocin-induced diabetic mice was significantly reduced from 15.81 ± 1.71 mmol L(-1) to 8.10 ± 0.88 mmol L(-1) after 10 d administration of the ß-carotene extract [100 mg kg(-1) body weight (BW)], and the increased food and water intakes in the diabetic mice were also significantly relieved after ß-carotene treatment. CONCLUSION: Our results suggested that extraction of ß-carotene from Spirulina platensis had potential prospects in scaled-up industrialisation and healthcare applications.

TCM network pharmacology: A new trend towards combining computational, experimental and clinical approaches.

Traditional Chinese medicine (TCM) is a precious treasure of the Chinese nation and has unique advantages in the prevention and treatment of diseases. The holistic view of TCM coincides with the new generation of medical research paradigm characterized by network and system. TCM gave birth to a new method featuring holistic and systematic "network target", a core theory and method of network pharmacology. TCM is also an important research object of network pharmacology. TCM network pharmacology, which aims to understand the network-based biological basis of complex diseases, TCM syndromes and herb treatments, plays a critical role in the origin and development process of network pharmacology. This review introduces new progresses of TCM network pharmacology in recent years, including predicting herb targets, understanding biological foundation of diseases and syndromes, network regulation mechanisms of herbal formulae, and identifying disease and syndrome biomarkers based on biological network. These studies show a trend of combining computational, experimental and clinical approaches, which is a promising direction of TCM network pharmacology research in the future. Considering that TCM network pharmacology is still a young research field, it is necessary to further standardize the research process and evaluation indicators to promote its healthy development.

Mixed fermentation of Spirulina platensis with Lactobacillus plantarum and Bacillus subtilis by random-centroid optimization.

Spirulina platensis is a high-nutrient blue-green alga with a long history as a food supplement. In this study, the mixed fermentation of Spirulina with Lactobacillus plantarum and Bacillus subtilis was investigated using random-centroid optimization to improve deodorization of off-flavor and hydrolysis of protein. Fermented Spirulina with the maximum total viable counts of both organisms achieved best sensory characteristics and degree of proteolysis among those with different maximum biomass. The mixed fermentation noticeably reduced the volatile compounds of Spirulina, and yielded the highest relative contents (>85%) of acetoin and other odorants collectively producing a creamy aroma. Approximately one-third of the Spirulina proteins were hydrolyzed, yielding over 16% polypeptides and increasing the ratio of essential amino acids to total free amino acids to 1.5-fold compared with unfermented Spirulina. Fermentation with B. subtilis and L. plantarum effectively improved the odor and protein availability of Spirulina.

[The effects of anthocyanin on chronic inflammatory pain induced by complete Freund's adjuvant and its mechanism].

OBJECTIVE: To investigate the effects and mechanisms of anthocyanin from Ligustrum vicaryi on chronic inflammatory pain induced by complete Freund's adjuvant. METHODS: Thirty male SD rats were randomly divided into three groups (n=10):normal saline control group (NS), chronic inflammatory pain model group(Mod, injected with complete Freund's adjuvant(CFA) 100 µl to the left hind leg), anthocyanin treatment group(Ant, dosed with anthocyanins (90 m), mechanical pain threshold (MPT), and left toe volume in each group were measured before modeling and 1,3,5,7,9,11,13 days after operation. Antioxidant indexes in serum were mensurated by spectrophotometer, and the total capsaicin receptor (TRPV1) and phosphorylated capsaicin receptor (p-TRPV1) in hippocampus were detected by Western blot. RESULTS: In comparison with controls, HPT and MPT were improved (P<0.05),toe swelling was reduced(P<0.05), the serum level of SOD was increased (P<0.01), while the levels of MDA and NO were decreased (P<0.05), the ratio of P-TRPV1/TRPV1 protein was depressed in Mod rat hippocampal region treated with anthocyanin. CONCLUSIONS: The results show that anthocyanins has an analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism may be related to the improvement of antioxidant capacity and the reduction of TRPV1 phosphorylation.